title: Contraction and intracellular calcium-ion elevation of cultured human aortic smooth muscle cells by endothelin-1, vasoactive intestinal contractor (VIC) and the derivatives.
abstract: Effects of endothelin (ET) family peptides and their derivatives on cellular contraction and calcium-ion level were examined by using cultured human vascular smooth muscle cells (VSM). Contraction of cultured human VSM, isolated from human fetal aortic segments, was induced within 1 min after the treatment with ET-1 (100 nM) as seen in the changes of cytosolic calcium-ion localization. In parallel with the cell contraction, cytosolic calcium-ion level in the human VSM increased very rapidly and then dropped with some oscillation as determined by Anchorage Cell Analyzing System. It was noted that transient calcium-ion mobilization rather than sustained calcium-ion influx was significant in the contraction of cultured human VSM. Vasoactive intestinal contractor (VIC), three amino acids different from ET-1, had less activity in increase of intracellular calcium-ion level and in percent of response cells than ET-1, ET-2, and VIC-S4L6 (one amino acid different from ET-1). EC50 of ET-1, VIC-S4L6, ET-2, and VIC were 0.5 nM, 0.6 nM, 2.0 nM, and 20 nM, respectively. VIC-like peptide (VIC-LP), 16 amino acids fragment of VIC precursor protein, had no effect with a single administration of up to 10 microM. However, the increase in calcium-ion level by VIC was suppressed with a prior treatment of cells with high concentration (10 microM) of VIC-LP. The establishment of cultured human VSM for the simultaneous examination of the contraction and calcium-ion level will provide a new system to study signal transduction of vasocontractor peptides.
authors.affiliation: National Institute of Bioscience and Human-Technology, Tsukuba, Ibaraki, Japan.
authors.forename: A
authors.initials: A
authors.lastname: Iwashima
authors.forename: M
authors.initials: M
authors.lastname: Kobayashi
authors.forename: K
authors.initials: K
authors.lastname: Saida
authors.forename: H
authors.initials: H
authors.lastname: Kagamu
authors.forename: S
authors.initials: S
authors.lastname: Ohashi
authors.forename: M
authors.initials: M
authors.lastname: Arakawa
authors.forename: Y
authors.initials: Y
authors.lastname: Mitsui
mesh_terms: D000595:Amino Acid Sequence; D001011:Aorta; D002118:Calcium / Q000378:metabolism*; D002478:Cells, Cultured; D004305:Dose-Response Relationship, Drug; D004622:Embryo, Mammalian; D019332:Endothelin-1 / Q000008:administration & dosage / Q000494:pharmacology*; D006801:Humans; D036341:Intercellular Signaling Peptides and Proteins; D008297:Male; D008969:Molecular Sequence Data; D009119:Muscle Contraction; D009131:Muscle, Smooth, Vascular / Q000187:drug effects* / Q000502:physiology*; D010455:Peptides / Q000008:administration & dosage / Q000494:pharmacology*; D017385:Sequence Homology
chemical_list: D019332:Endothelin-1; D036341:Intercellular Signaling Peptides and Proteins; D010455:Peptides; C059568:vasoactive intestinal constrictor; D002118:Calcium